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Bartholomew, Rachel A.

Publications and source records attributed to Bartholomew, Rachel A..

At least 19 records

Emergency Management of Tomorrow Research: Emergency Operations Center of the Future Madison, WI Tabletop Exercise

As part of the Emergency Management (EM) of Tomorrow Research (EMOTR) program, sponsored by the Department of Homeland Security (DHS) Science and Technology (S&T) Directorate, Pacific Northwest National Laboratory (PNNL) developed concepts for the Emergency Operations Center (EOC) of the Future to provide recommendations to assist DHS S&T in future decision-making with regards to research and development (R&D) and investments toward establishing a framework for a national, coordinated approach to EM. PNNL is conducting tabletop exercises (TTXs) designed to assess the impacts and benefits of emerging technologies on EM organizations.

99 GENERAL AND MISCELLANEOUS↗

Emergency Management of Tomorrow Research – Task 3B Research and Development Community Awareness: Eliciting Emergency Management Stakeholder Input

The Department of Homeland Security (DHS) Science and Technology Directorate (S&T) is partnering with Pacific Northwest National Laboratory (PNNL) to execute the Emergency Management (EM) of Tomorrow Research (EMOTR) program to identify current EM research, elicit capability needs from EM practitioners, and identify where technology, such as artificial intelligence (AI), may benefit the future of EM and emergency operations centers. This report details the methodology, analysis, and insights of interviews and focus groups conducted as part of the task to elicit stakeholder input.

99 GENERAL AND MISCELLANEOUS↗

End-User Perspectives on Using Quantitative Real-Time PCR and Genomic Sequencing in the Field

Quantitative real-time PCR and genomic sequencing have become mainstays for performing molecular detection of biological threat agents in the field. There are notional assessments of the benefits, disadvantages, and challenges that each of these technologies offers according to findings in the literature. However, direct comparison between these two technologies in the context of field-forward operations is lacking. Most market surveys, whether published in print form or provided online, are directed to product manufacturers who can address their respective specifications and operations. One method for comparing these technologies is surveying end-users who are best suited for discussing operational capabilities, as they have hands-on experience with state-of-the-art molecular detection platforms and protocols. These end-users include operators in military defense and first response, as well as various research scientists in the public sector such as government and service laboratories, private sector, and civil society such as academia and nonprofit organizations performing method development and executing these protocols in the field. Our objective was to initiate a survey specific to end-users and their feedback. We developed a questionnaire that asked respondents to (1) determine what technologies they currently use, (2) identify the settings where the technologies are used, whether lab-based or field-forward, and (3) rate the technologies according to a set list of criteria. Of particular interest are assessments of sensitivity, specificity, reproducibility, scalability, portability, and discovery power. This article summarizes the findings from the end-user perspective, highlighting technical and operational challenges.

59 BASIC BIOLOGICAL SCIENCES↗

Making sense of the 2018 National Biodefense Strategy

A tool dubbed the “spaghetti monster” helps users navigate the tangle of laws, directives, and agencies that make up the US biodefense enterprise—and reveals its formidable complexity. It has been 100 years since the end of World War I, the “war to end all wars,” which set the precedent for modern biological warfare. The 1925 Geneva Protocol subsequently banned the use of biological weapons in war but did not stop the proliferation of state-sponsored bioweapons programs. With the spread of such programs and other biologically based threats, the United States and other nations have made a variety of efforts to defend against them.

99 GENERAL AND MISCELLANEOUS↗

A Publicly Available Landscape Analysis Tool for Biodefense Policy

Outbreaks of infectious diseases, coupled with an increasing biological threat from non-state actors, highlight the continued need to prioritize biodefense. The Blue Ribbon Panel on Biodefense has noted that the US remains ‘underprepared’ for a catastrophic biological attack or pandemic and emphasized increased coordination is essential. Consistent with this recommendation, the FY17 National Defense Authorization Act tasked the Secretaries of Defense, Health and Human Services, Homeland Security, and Agriculture to “jointly develop a national biodefense strategy and associated implementation plan, which shall include a review and assessment of biodefense policies, practices, programs and initiatives.”

Omberg, Kristin M.↗

Cyber Deterrence and Stability

Throughout the 20th and early 21st centuries, deterrence and arms control have been cornerstones of strategic stability between the superpowers. However, the weaponization of the cyber realm by State actors and the multipolar nature of cyber conflict now undermines that stability. Strategic stability is the state in which nations believe that if they act aggressively to undermine U.S. national interests and the post-World War II liberal democratic order, the consequences will outweigh the benefits. The sense of lawlessness and lack of consequences in the cyber realm embolden States to be more aggressive in taking actions that undermine stability. Accordingly, this paper examines 1) the role of deterrence and arms control in securing cyber stability, and 2) the limitations and challenges associated with these traditional national security paradigms as applied to this emerging threat domain. This paper demonstrates that many 20th-century deterrence and arms control concepts are not particularly applicable in the cyber realm. However, they are not entirely irrelevant. The United States can distill lessons learned from this rich deterrence and arms control experience to develop and deploy a strategy to advance cyber stability.

97 MATHEMATICS AND COMPUTING↗

Defining the Synthetic Biology Supply Chain

In this article, a team of experts in synthetic biology, data analytics, and national security describe the overall supply chain surrounding synthetic biology. The team analyzes selected interactions within that network to better understand the risks raised by synthetic biology and identifies opportunities for risk mitigation. To introduce the concept, the article will briefly describe how an understanding of supply chains has been important in promoting nuclear nonproliferation objectives. The article concludes by assessing the structure and networks identified in the supply chains to reveal potential opportunities for future biodefense research and development; options for additional information exchange; and means to interdict, detect, or deter suspicious activity.

nuclear supply chain↗

Evaluation of PCR Systems for Field Screening of Bacillus anthracis

There is little published data on the performance of hand-portable polymerase chain reaction (PCR) instruments that could be used by first responders to determine if a suspicious powder contains a potential biothreat agent. We evaluated five commercially available hand-portable PCR instruments for detection of Bacillus anthracis (Ba). We designed a cost-effective, statistically-based test plan that allows instruments to be evaluated at performance levels ranging from 0.85-0.95 lower confidence bound (LCB) on the probability of detection (POD) at confidence levels of 80-95%. We assessed specificity using purified genomic DNA from 13 Ba strains and 18 Bacillus near neighbors, interference with 22 common hoax powders encountered in the field, and PCR inhibition when Ba spores were spiked into these powders. Our results indicated that three of the five instruments achieved >0.95 LCB on the POD with 95% confidence at test concentrations of 2,000 genome equivalents/mL (comparable to 2,000 spores/mL), displaying more than sufficient sensitivity for screening suspicious powders. These instruments exhibited no false positive results or PCR inhibition with common hoax powders, and reliably detected Ba spores spiked into common hoax powders, though some issues with instrument controls were observed. Our testing approach enables efficient instrument performance testing to a statistically rigorous and cost-effective test plan to generate performance data that will allow users to make informed decisions regarding the purchase and use of biodetection equipment in the field.

59 BASIC BIOLOGICAL SCIENCES↗

Evaluation of Immunoassays and General Biological Indicator Tests for Field Screening of Bacillus anthracis and Ricin

The goal of this testing was to evaluate the ability of currently available commercial off-the-shelf (COTS) biological indicator tests and immunoassays to detect Bacillus anthracis (Ba) spores and ricin. In general, immunoassays provide more specific identification of biological threats as compared to indicator tests [3]. Many of these detection products are widely used by first responders and other end users. In most cases, performance data for these instruments are supplied directly from the manufacturer, but have not been verified by an external, independent assessment [1]. Our test plan modules included assessments of inclusivity (ability to generate true positive results), commonly encountered hoax powders (which can cause potential interferences or false positives), and estimation of limit of detection (LOD) (sensitivity) testing.

59 BASIC BIOLOGICAL SCIENCES↗

Field application of pathogen detection technologies

Over the last 10 years there has been a significant increase in commercial products designed for field-based detection of microbial pathogens. This is due, in part, to the anthrax attacks in the United States in 2001, and the need for first responders to quickly identify the composition of suspected white powders and other potential biothreats. Demand for rapid detection is also driven by the need to ensure safe food, water, and environmental systems. From a technology perspective, rapid identification methods have largely capitalized on PCR and other molecular recognition techniques that can be deployed as robust field instrumentation. Examples of the relevant needs include the ability to: 1) declare a water distribution system free of microbial pathogens after a pipe/main break repair; 2) assess risks of contamination such as when produce production and processing plants are located near concentrated animal feeing operations; 3) evaluate the safety of ready-to-eat products; 4) determine the extent of potential serious disease outbreaks in remote and/or disaster stricken areas where access to clinical laboratories is not an immediate option; and 5) quickly assess credible biological terrorism events. Many of the principles underlying rapid detection methods are derived from methods for environmental microbiology, but there is a dearth of literature describing and evaluating field-based detection systems. Thus, the aims of this chapter are to: 1) summarize the different kinds of commercially available sampling kits and field-based biological detectors; 2) highlight some of the continued challenges of sample preparation to stimulate new research towards minimizing the impact of inhibitors on PCR-based detection systems; 3) describe our general rationale and statistically-based approach for instrument evaluation; 4) provide statistical and spatial guidelines for developing valid sampling plans; and 5) summarize some current needs and emerging technologies. This information is presented both to highlight the state of the field, and to also highlight major questions that students may wish to consider investigating further. Where possible we will cite studies that have been conducted and published either in traditional peer-reviewed or other literature (e.g., AOAC International Methods).

Straub, Tim M.↗

National Laboratory Support for First Responders’ Biodetection Needs

First responders know that white powder scenarios – or suspected biological threats – require quick and decisive action. Having the right field equipment available to identify suspicious substances can be complicated, challenging, and expensive. With support from the Department of Homeland Security (DHS) Science and Technology Directorate (S&T), Pacific Northwest National Laboratory (PNNL) has been working with the first responder community to identify biodetection technology and information needs and gaps, and transition solutions to the first responder community. PNNL’s “ground-up approach” involves first responders and stakeholders early in the process and culminates in the transition of information and knowledge to improve biological response capabilities.

biodetection↗

Contributions of the [NiFe]- and [FeFe]-hydrogenase to H2 production in Shewanella oneidensis MR-1 as revealed by isotope ratio analysis of evolved H2

Shewanella oneidensis MR-1 encodes both a [NiFe]- and an [FeFe]-hydrogenase. While the output of these proteins has been characterized in mutant strains expressing only one of the enzymes, the contribution of each to H2 synthesis in the wild-type organism is not clear. Here we use stable isotope analysis of H2 in the culture headspace, along with transcription data and measurements of the concentrations of gases in the headspace, to characterize H2 production in the wild-type strain. After most of the O2 in the headspace had been consumed, H2 was produced and then consumed by the bidirectional [NiFe]-hydrogenase. Once the cultures were completely anaerobic, a new burst of H2 synthesis catalyzed by both enzymes took place. Our data is consistent with the hypothesis that at this point in the culture cycle, a pool of electrons is shunted toward both hydrogenases in the wild-type organism, but that in the absence of one of the hydrogenases, the flux is redirected to the available enzyme. To our knowledge, this is the first use of stable isotope analysis of a metabolic product to elucidate substrate flux through two alternative enzymes in the same cellular system.

stable isotope, bacteria, metabolism, anaerobic, e↗

Biodetection Technologies for First Responders: 2014 Edition

This report summarizes commercially-available, hand-portable technologies that can be used by first responders in the field. This is not meant to be an exhaustive list, nor an endorsement of any technology described herein. Rather, this report is meant to provide useful information about available technologies to help end-users make informed decisions about biodetection technology procurement and use. Information listed in this report is primarily vendor-provided; however, where possible it has been supplemented with additional information obtained from publications, reports, and websites. Manufacturers were given the chance to review summaries of their technologies from August through November 2013 to verify the accuracy of technical specifications, available references, and pricing.

biodefense↗

BioCat 2.0

The U.S. Department of Homeland Security (DHS) National Biosurveillance Integration Center (NBIC) was established in 2008 with a primary mission to “(1) enhance the capability of the Federal Government to (A) rapidly identify, characterize, localize, and track a biological event of national concern by integrating and analyzing data relating to human health, animal, plant, food, and environmental monitoring systems (both national and international); and (B) disseminate alerts and other information to Member Agencies and, in coordination with (and where possible through) Member Agencies, to agencies of State, local, and tribal governments, as appropriate, to enhance the ability of such agencies to respond to a biological event of national concern; and (2) oversee development and operation of the National Biosurveillance Integration System (NBIS).” Inherent in its mission then and the broader NBIS, NBIC is concerned with the identification, understanding, and use of a variety of biosurveillance models and systems. The goal of this project is to characterize, evaluate, classify, and catalog existing disease forecast and prediction models that could provide operational decision support for recognizing a biological event having a potentially significant impact. Additionally, gaps should be identified and recommendations made on using disease models in an operational environment to support real-time decision making.

biosurveillance↗

Evaluation of the FilmArray® system for detection of Bacillus anthracis, Francisella tularensis, and Yersinia pestis

To evaluate the sensitivity and specificity of the Idaho Technologies FilmArray® Biothreat Panel for the detection of Bacillus anthracis (Ba), Francisella tularensis (Ft), and Yersinia pestis (Yp) DNA, and demonstrate the detection of Ba spores. Methods and Results: DNA samples from Ba, Ft and Yp strains and near-neighbors, and live Ba spores were analyzed using the Biothreat Panel, a multiplexed PCR-based assay for 17 pathogens and toxins. Sensitivity studies with DNA suggest a limit of detection of 250 genome equivalents (GEs) per sample. Furthermore, the correct call of Ft, Yp or Bacillus species was made in 63 of 72 samples tested at 25 GE or less. With samples containing 25 Ba Sterne spores, at least one of the two possible Ba markers were identified in all samples tested. We observed no cross-reactivity with near-neighbor DNAs.

Biodetection, PCR, hand portable, pathogens, biote↗

Defining cell culture conditions to improve human norovirus infectivity assays

Significant difficulties remain for determining whether human noroviruses (hNoV) recovered from water, food, and environmental samples are infectious. Three-dimensional tissue culture of human intestinal cells has shown promise in developing an infectivity assay, but reproducibility, even within a single laboratory, remains problematic. From the literature and our observations, we hypothesized that the common factors that leads to more reproducible hNoV infectivity in vitro requires that the cell line be 1) of human gastrointestinal origin, 2) expresses apical microvilli, and 3) be a positive secretor cell line. The C 2 BBe1 cell line, which is a brush-border producing clone of Caco-2, meets these three criteria. When challenged with Genogroup II viruses, we observed a 2 Log10 increase in viral RNA titer. A passage experiment with GII viruses showed evidence of the ability to propagate hNoV by both reverse transcription quantitative PCR (qRT-PCR) and microscopy. Using 3-D C 2 BBe1 cells improves reproducibility of the infectivity assay for hNoV, but the assay can still be variable. Two sources of variability include the cells themselves (mixed phenotypes of small and large intestine) and initial titer measurements using quantitative reverse transcription PCR (qRT-PCR) that measures all RNA vs. plaque assays that measure infectious virus.

C2BBe1 cells↗